Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.27.4 (ribonuclease)
 6,621 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

GnRH regulates gonadotropin biosynthesis and release in the anterior pituitary via specific receptors. Although extrapituitary expression and action of GnRH have been shown in some species, in the human it is not clear whether GnRH has a peripheral action. In this study we sought to determine whether the human ovary expresses GnRH receptor (GnRHR) messenger ribonucleic acid (mRNA). Ovarian tissues from 11 women (32-61 yr old) and granulosa-lutein (GL) cells purified from follicular aspirates of 51 women undergoing oocyte retrieval for in vitro fertilization were analyzed by ribonuclease protection assay and reverse transcriptase-polymerase chain reaction (RT-PCR). Human pituitaries, lymphocytes, and placenta were also studied. Measurable levels of GnRHR mRNA were found by ribonuclease protection assay in 2 of 10 ovaries, in 2 of 4 GL cells preparations from women whose ovarian hyperstimulation involved a GnRH agonist, in GL cells from 3 women whose ovarian hyperstimulation involved a GnRH antagonist, and in human pituitaries. Relative to the total amount of RNA analyzed, the level of GnRHR mRNA was about 200-fold lower in the ovary than in the pituitary. A sequence of 314 basepairs of GnRHR mRNA was amplified by RT-PCR in the pituitary, in 9 of 10 ovaries, and in 4 of 5 GL cell preparations. No message could be amplified in human lymphocytes, and placental specimens showed a weak signal. The relative GnRHR mRNA levels in GL cells from 13 women analyzed by quantitative RT-PCR showed a wide range of individual differences. These results suggest that GnRHR mRNA is expressed in GL cells and the human ovary across different functional stages, implying that multiple ovarian compartments may express GnRH receptors. The administration of GnRH analogs may have a further direct action on the human ovary.
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PMID:Gonadotropin-releasing hormone receptor gene expression in human ovary and granulosa-lutein cells. 785 1

Prolonged exposure of the GnRH receptor to high concentrations of GnRH leads to receptor down-regulation. The role of altered receptor biosynthesis in this agonist-induced receptor down-regulation was investigated in the mouse gonadotrope cell line, alpha T3-1 cells. After exposure to 1 microM GnRH for 24 h, the number of GnRH receptor-binding sites in alpha T3-1 cells decreased to 25 +/- 6% of the control levels. No corresponding changes were observed in GnRH receptor messenger RNA (mRNA) using either quantitative ribonuclease protection/solution hybridization assay or Northern blot analysis. However, when the ability of this RNA to direct the synthesis of functional GnRH receptors was examined by quantitative assessment of the voltage clamp response in Xenopus oocytes, GnRH-induced currents in oocytes injected with RNA isolated from down-regulated cells was reduced to 40 +/- 13% of the response obtained after the injection of RNA from control alpha T3-1 cells. Thus, although GnRH receptor mRNA levels were not altered, the ability of cellular RNA isolated from alpha T3-1 cells to direct the synthesis of functional GnRH receptors was regulated in concert with receptor binding. To investigate the possibility that GnRH receptor mRNA translational efficiency was reduced, the distribution of polyribosome-associated GnRH receptor mRNA was studied. Polyribosome-associated mRNA was separated by linear sucrose gradient, and GnRH receptor mRNA distribution was determined by ribonuclease protection assay. GnRH receptor mRNA distribution shifted from the largest to smaller polyribosome and monosome fractions in cells exposed to GnRH compared to controls. The weighted mean of GnRH receptor mRNA distribution among eight fractions shifted from fraction 5.924 +/- 0.06 in control polysomes to fraction 5.45 +/- 0.219 for polysomes from down-regulated cells (P < 0.05). These results demonstrate that GnRH receptor down-regulation is accompanied by decreased GnRH receptor mRNA translation in the absence of any change in GnRH receptor mRNA levels. These data suggest that decreased efficiency of GnRH receptor mRNA translation contributes to the down-regulation of this receptor in alpha T3-1 cells.
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PMID:Translational regulation of the gonadotropin-releasing hormone receptor in alpha T3-1 cells. 786 66

The GnRH receptor (GnRH-R) is a cell surface, G protein-coupled receptor that is highly expressed in pituitary gonadotropes. Activation of the receptor by GnRH stimulates the release of FSH and LH. Pituitary GnRH-R numbers and, hence, gonadotrope responsiveness to GnRH vary under different conditions and are regulated to a large extent by GnRH itself. To study the transcriptional regulation of the GnRH-R gene, a genomic clone containing 1.2 kilobases (kb) of the 5'-flanking region of mouse GnRH-R gene was isolated and characterized. A major transcriptional start site was identified 62 nucleotides upstream of the translational start site by primer extension and ribonuclease protection analyses. The promoter region does not contain canonical TATA sequences in the appropriate location. To determine whether this putative promoter is functional, it was subcloned into a luciferase reporter plasmid (GnRH-RLuc), and its transient expression was studied in cell lines of gonadotrope (alpha T3-1) and somatolactotrope (GH3) origins as well as those of nonpituitary origin (JEG-3 and CV-1). Luciferase activity was increased in alpha T3-1 (246-fold +/- 34.5-fold; P < 0.005) compared with the promoterless vector control but was considerably lower in GH3 (41-fold +/- 3.9-fold; P < 0.005), JEG-3 (12-fold +/- 0.9-fold; P < 0.005) and CV-1 (8-fold +/- 1.3-fold) indicating that GnRH-RLuc is preferentially expressed in cells of gonadotrope origin. Furthermore, GnRH agonist stimulated luciferase activity 3.4-fold +/- 0.3-fold (P < 0.005) above basal levels in GH3 cells cotransfected with rat GnRH-R complementary DNA, indicating that the GnRH-R promoter sequence is responsive to this ligand. In summary, we have identified and partially characterized the promoter region of the mouse GnRH-R and demonstrated that the regulatory elements for tissue-specific expression as well as for GnRH regulation are present within a 1.2-kb 5'-flanking region of the mouse GnRH-R gene.
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PMID:Isolation and characterization of the 5'-flanking region of the mouse gonadotropin-releasing hormone receptor gene. 798 11

In rats, FSHbeta gene expression and FSH secretion are increased and decreased, respectively, by pituitary activin and follistatin. Because little information is available on the paracrine control of FSH secretion in the primate, follistatin and activin/inhibin beta(B) messenger RNA (mRNA) levels were measured in pituitaries of adult male rhesus monkeys 6 weeks after castration or sham surgery (n = 5/group). Follistatin mRNA was determined by quantitative RT-PCR assay using oligonucleotide primers designed to span exons 3-5 of the human follistatin gene. Activin/inhibin beta(B) mRNA levels were measured by ribonuclease protection. Orchidectomy resulted in a 100-fold increase in plasma FSH concentrations and a 60-fold rise in those of LH. In castrated monkeys, levels of mRNA encoding FSHbeta, LHbeta, alpha- subunit, and GnRH receptor (GnRH-R) were increased 21-, 2.1-, 1.7-, and 1.7-fold, respectively (P < 0.01). Levels of pituitary follistatin and activin/inhibin beta(B) mRNAs, however, were similar in castrated and intact animals. These data suggest that the paracrine control of FSH secretion in the male differs substantially in primates and rodents. Specifically, the relatively greater postcastration rise in FSHbeta gene expression and FSH secretion in the adult male monkey may result because in this species pituitary follistatin gene expression does not increase after orchidectomy, as it does in the rat.
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PMID:Pituitary follistatin and activin gene expression, and the testicular regulation of FSH in the adult Rhesus monkey (Macaca mulatta). 1141 6